Kinase and Phosphatase Mechanism Analysis Service Based on SPR Technology

Protein phosphorylation is a post-translational modification that regulates numerous cellular physiological processes and regulates a variety of intracellular signalling pathways. Protein phosphorylation is a reversible dynamic process regulated by the competing activities of protein kinases and phosphatases.

• Protein kinase

Protein kinases hydrolyse adenosine triphosphate (ATP) and transfer the terminal phosphate group (PO4) of ATP to the hydroxyl groups of various amino acid residues, thereby modifying the protein from hydrophobic to hydrophilic. The phosphorylated amino acids can interact with other proteins and thus assemble and de-assemble the protein complex. Each protein kinase has a similar three-dimensional catalytic structural domain. As shown in the diagram below, this 3D catalytic structural domain consists of 250-300 amino acids and includes 2 subdomains. These two sub-domains are the N-terminal and C-terminal, which are connected by a peptide scaffold and form a deep groove between them, allowing binding to peptide substrates and ATP molecules. Protein kinases exert their action by phosphorylating their respective downstream specific protein substrates. Dysregulation of protein kinases leads to changes in enzyme activity, enzyme expression, subcellular localisation, enzyme stability, protein-protein interactions and more. Aberrant regulation of kinase-mediated signalling pathways is responsible for all major human pathological conditions, such as cancer, cardiovascular disease, neurodegenerative diseases and metabolic disorders.

Fig.1 Schematic representation of the structure of the catalytic subunit of protein kinase A. (N-terminal is light blue, C-terminal is red, peptide substrate is yellow and ATP is orange)(Kobe, B.;et al, 2005)

• Protein kinase/phosphatase regulated signalling pathways

Phosphatases function in the opposite way to protein kinases, removing phosphate groups from phosphorylated proteins by hydrolysing the phosphate monoester into a phosphate group and a molecule with a free hydroxyl group. Dephosphorylation occurs mainly on serine, threonine and tyrosine residues.

Fig.2 Schematic diagram of protein phosphorylation process (Ardito, F.;et al, 2017)

As shown above, phosphorylation and dephosphorylation are often the switches that activate key regulatory proteins and control signalling pathways. Once the phosphorylation and dephosphorylation processes become abnormal, the associated signalling pathways become dysfunctional. As a result, abnormal protein phosphorylation has been implicated in the development of a wide range of diseases, from cancer to inflammatory diseases, diabetes, infectious diseases, cardiovascular diseases and more. A large number of protein kinases and phosphatases have now become popular targets for drug development.

Surface plasmon resonance (SPR) in kinase and phosphatase mechanism analysis

Since phosphorylation is a reversible process, phosphorylation and dephosphorylation is an important molecular switch. The phosphorylation and dephosphorylation processes of different functional proteins control different signalling pathways in the living organism and are relevant to normal life activities. Therefore, it is essential to study the mechanism of protein phosphorylation, which is important for the development of most disease-related drugs. In the study of the mechanism of protein phosphorylation, the analysis of protein-related interactions is an important step in explaining the mechanism at the molecular level. If you have research needs in this area, consider our technology platform. SPR is a technology that provides high throughput and high sensitivity molecular interaction analysis services. The diagram below illustrates the general workflow of our SPR technology platform.

Fig.3 BIAchip™ in the process of kinase and phosphatase mechanism analysis

The SPR technology platform can be used for both mechanistic studies and drug screening in the context of protein phosphorylation research. Compared to traditional molecular interaction analysis methods, SPR technology has higher throughput, higher sensitivity and more accurate results.

• Phosphorylation mechanisms of different proteins

Our SPR technology platform provides you with high-throughput analysis of protein-related molecular interactions, helping you to explain the underlying mechanisms at the molecular level and laying a solid foundation for subsequent downstream studies. The high-throughput technology platform ensures that your relevant studies are reproducible, saving you time. In addition, based on the high flexibility of our SPR technology platform, the chip can be customised for any design and idea. After placing your order, one of our expert chip designers will contact you and you will receive feedback within 2-3 days after we understanding your requirements.

• New drug development for protein phosphorylation-related diseases

The switching function of protein phosphorylation not only makes it a source of disease, but also a potential drug target. The high-throughput nature of SPR technology is a great advantage in the drug screening process and can drive the entire drug development process. There is no more time and cost effective option than SPR technology.

Choosing SPR technology platform of Creative Proteomics, you will greatly save time and money costs owing to high-throughput intermolecular interaction detection. All services are available on a 24/7/365 basis. If you have any questions or suggestions about our SPR platform, please feel free to contact us right now.

For research use only. Not intended for any clinical use.